Synthesis of the 2-Amino-4-phosphonobutanoic Acid Analogues (E)- and (Z)-2-Amino-2,3-methano-4-phosphonobutanoic Acid and Their Evaluation as Inhibitors of Hippocampal Excitatory Neurotransmission

The cyclopropyl compounds (Z)- and (E)-2-amino-2,3-methano-4-phosphonobutanoic acid, 5 and 6, respectively, were prepared as constrained analogues of 2-amino-4-phosphonobutanoic acid (AP4), a selective glutamate receptor ligand. A Homer-Emmons reaction of trimethyl N-(benzyloxycarbonyl)phosphonoglycinate with 2-(diethoxy- phosphinyl)acetaldehyde gave the protected dehydroamino acids 9 and 10, which were individually subjected to the following sequence of reactions: cycloaddition of diazomethane, photoelimination of N₂, and acid hydrolysis, to give 5 and 6, respectively. Extracellular recording techniques were used to evaluate the abilities of 5 and 6 to block evoked synaptic transmission in specific neuronal pathways of the rat hippocampal slice. In the lateral perforant path (LPP) 5 and 6 were equipotent and possessed IC₅₀ values of 18 and 17 μM, respectively. In the medial perforant path (MPP), 6 (IC₅₀ = 81 μM) was much more potent than 5 (IC₅₀ = 1580 μM). In paired pulse experiments which differentiate presynaptic and postsynaptic inhibition, 5 and 6 enhanced the second response to the same extent as L-AP4, suggesting a presynaptic site of action for these compounds. In contrast, the cyclopentyl AP4 analogues 3 and 4 enhanced the second response to a lesser extent. It was concluded that the biologically active conformation of AP4 in the LPP is different than in the MPP. In order to explain the same potency of 5 and 6 in the LPP, it was postulated that the two analogues assume a conformation that allows their functional groups to occupy the same relative place in space. Molecular modeling showed that the best overlap was achieved when the αC–αC-γC-P dihedral angle for 5 was in the range of 130° to 180° and that of 6 was in the range of -130° to -180°. The results suggest that the bioactive conformation of AP4 in the LPP is an extended one.